Stepwise investigation from Ago2-RIP assay demonstrated that Anti-Ago2 antibody markedly enriched the SLC2A1-AS1 and miR-378a-3p in ESCC cells, notably, when miR-378a-3p was overexpressed, the enrichment of SLC2A1-AS1 was dramatically enhanced, suggesting that SLC2A1-AS1 and miR-378a-3p appeared in the same RISC in ESCC cells. Here, AGO2 is linked to esophageal squamous cell carcinoma.